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<title xml:lang="en">High infection rate of zoonotic
<italic>Eucoleus aerophilus</italic>
infection in foxes from Serbia</title>
<author>
<name sortKey="Lalosevi, Vesna" sort="Lalosevi, Vesna" uniqKey="Lalosevi V" first="Vesna" last="Laloševi">Vesna Laloševi</name>
<affiliation>
<nlm:aff id="AFF1">
<addr-line> Faculty of Agriculture, University of Novi Sad Trg Dositeja Obradovica 8 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Lalosevi, Dusan" sort="Lalosevi, Dusan" uniqKey="Lalosevi D" first="Dušan" last="Laloševi">Dušan Laloševi</name>
<affiliation>
<nlm:aff id="AFF2">
<addr-line> Faculty of Medicine, University of Novi Sad Hajduk Veljkova 3 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
<affiliation>
<nlm:aff id="AFF3">
<addr-line> Pasteur Institute Hajduk Veljkova 1 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey=" Apo, Ivan" sort=" Apo, Ivan" uniqKey=" Apo I" first="Ivan" last=" Apo">Ivan Apo</name>
<affiliation>
<nlm:aff id="AFF2">
<addr-line> Faculty of Medicine, University of Novi Sad Hajduk Veljkova 3 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Simin, Verica" sort="Simin, Verica" uniqKey="Simin V" first="Verica" last="Simin">Verica Simin</name>
<affiliation>
<nlm:aff id="AFF3">
<addr-line> Pasteur Institute Hajduk Veljkova 1 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Galfi, Annamaria" sort="Galfi, Annamaria" uniqKey="Galfi A" first="Annamaria" last="Galfi">Annamaria Galfi</name>
<affiliation>
<nlm:aff id="AFF1">
<addr-line> Faculty of Agriculture, University of Novi Sad Trg Dositeja Obradovica 8 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Traversa, Donato" sort="Traversa, Donato" uniqKey="Traversa D" first="Donato" last="Traversa">Donato Traversa</name>
<affiliation>
<nlm:aff id="AFF4">
<addr-line> Department of Comparative Biomedical Sciences, University of Teramo Teramo Italy</addr-line>
</nlm:aff>
</affiliation>
</author>
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<title xml:lang="en" level="a" type="main">High infection rate of zoonotic
<italic>Eucoleus aerophilus</italic>
infection in foxes from Serbia</title>
<author>
<name sortKey="Lalosevi, Vesna" sort="Lalosevi, Vesna" uniqKey="Lalosevi V" first="Vesna" last="Laloševi">Vesna Laloševi</name>
<affiliation>
<nlm:aff id="AFF1">
<addr-line> Faculty of Agriculture, University of Novi Sad Trg Dositeja Obradovica 8 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Lalosevi, Dusan" sort="Lalosevi, Dusan" uniqKey="Lalosevi D" first="Dušan" last="Laloševi">Dušan Laloševi</name>
<affiliation>
<nlm:aff id="AFF2">
<addr-line> Faculty of Medicine, University of Novi Sad Hajduk Veljkova 3 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
<affiliation>
<nlm:aff id="AFF3">
<addr-line> Pasteur Institute Hajduk Veljkova 1 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey=" Apo, Ivan" sort=" Apo, Ivan" uniqKey=" Apo I" first="Ivan" last=" Apo">Ivan Apo</name>
<affiliation>
<nlm:aff id="AFF2">
<addr-line> Faculty of Medicine, University of Novi Sad Hajduk Veljkova 3 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Simin, Verica" sort="Simin, Verica" uniqKey="Simin V" first="Verica" last="Simin">Verica Simin</name>
<affiliation>
<nlm:aff id="AFF3">
<addr-line> Pasteur Institute Hajduk Veljkova 1 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Galfi, Annamaria" sort="Galfi, Annamaria" uniqKey="Galfi A" first="Annamaria" last="Galfi">Annamaria Galfi</name>
<affiliation>
<nlm:aff id="AFF1">
<addr-line> Faculty of Agriculture, University of Novi Sad Trg Dositeja Obradovica 8 21000 Novi Sad Serbia</addr-line>
</nlm:aff>
</affiliation>
</author>
<author>
<name sortKey="Traversa, Donato" sort="Traversa, Donato" uniqKey="Traversa D" first="Donato" last="Traversa">Donato Traversa</name>
<affiliation>
<nlm:aff id="AFF4">
<addr-line> Department of Comparative Biomedical Sciences, University of Teramo Teramo Italy</addr-line>
</nlm:aff>
</affiliation>
</author>
</analytic>
<series>
<title level="j">Parasite</title>
<idno type="ISSN">1252-607X</idno>
<idno type="eISSN">1776-1042</idno>
<imprint>
<date when="2013">2013</date>
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<div type="abstract" xml:lang="en">
<p>The respiratory capillariid nematode
<italic>Eucoleus aerophilus</italic>
(Creplin, 1839) infects wild and domestic carnivores and, occasionally, humans. Thus far, a dozen of human infections have been published in the literature but it cannot be ruled out that lung capillariosis is underdiagnosed in human medicine. Also, the apparent spreading of
<italic>E. aerophilus</italic>
in different geographic areas spurs new studies on the epidemiology of this nematode. After the recognition of the first human case of
<italic>E. aerophilus</italic>
infection in Serbia, there is a significant merit in enhancing knowledge on the distribution of the nematode. In the present work the infection rate of pulmonary capillariosis was investigated in 70 red foxes (
<italic>Vulpes vulpes</italic>
) from the northern part of Serbia by autopsy. The estimated infection rate with
<italic>Eucoleus aerophilus</italic>
was 84%. In contrast, by copromicroscopic examination only 38% of foxes were positive. In addition, 10 foxes were investigated for the closely related species in nasal cavity,
<italic>Eucoleus boehmi</italic>
, and nine were positive. Our study demonstrates one of the highest infection rates of pulmonary capillariosis in foxes over the world.</p>
</div>
</front>
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<pmc article-type="research-article">
<pmc-dir>properties open_access</pmc-dir>
<front>
<journal-meta>
<journal-id journal-id-type="nlm-ta">Parasite</journal-id>
<journal-id journal-id-type="iso-abbrev">Parasite</journal-id>
<journal-id journal-id-type="publisher-id">parasite</journal-id>
<journal-title-group>
<journal-title>Parasite</journal-title>
</journal-title-group>
<issn pub-type="ppub">1252-607X</issn>
<issn pub-type="epub">1776-1042</issn>
<publisher>
<publisher-name>EDP Sciences</publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="pmid">23340229</article-id>
<article-id pub-id-type="pmc">3718516</article-id>
<article-id pub-id-type="doi">10.1051/parasite/2012003</article-id>
<article-id pub-id-type="publisher-id">parasite120020</article-id>
<article-id pub-id-type="other">10.1051/parasite/2012003</article-id>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Research Article</subject>
</subj-group>
</article-categories>
<title-group>
<article-title>High infection rate of zoonotic
<italic>Eucoleus aerophilus</italic>
infection in foxes from Serbia</article-title>
<trans-title-group xml:lang="fr">
<trans-title>Prévalence élevée d’infection zoonotique par
<italic>Eucoleus aerophilus</italic>
chez les renards de Serbie</trans-title>
</trans-title-group>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname>Lalošević</surname>
<given-names>Vesna</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
<xref ref-type="corresp" rid="FN1">*</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Lalošević</surname>
<given-names>Dušan</given-names>
</name>
<xref ref-type="aff" rid="AFF2">2</xref>
<xref ref-type="aff" rid="AFF3">3</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Čapo</surname>
<given-names>Ivan</given-names>
</name>
<xref ref-type="aff" rid="AFF2">2</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Simin</surname>
<given-names>Verica</given-names>
</name>
<xref ref-type="aff" rid="AFF3">3</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Galfi</surname>
<given-names>Annamaria</given-names>
</name>
<xref ref-type="aff" rid="AFF1">1</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Traversa</surname>
<given-names>Donato</given-names>
</name>
<xref ref-type="aff" rid="AFF4">4</xref>
</contrib>
</contrib-group>
<aff id="AFF1">
<label>1</label>
<addr-line> Faculty of Agriculture, University of Novi Sad Trg Dositeja Obradovica 8 21000 Novi Sad Serbia</addr-line>
</aff>
<aff id="AFF2">
<label>2</label>
<addr-line> Faculty of Medicine, University of Novi Sad Hajduk Veljkova 3 21000 Novi Sad Serbia</addr-line>
</aff>
<aff id="AFF3">
<label>3</label>
<addr-line> Pasteur Institute Hajduk Veljkova 1 21000 Novi Sad Serbia</addr-line>
</aff>
<aff id="AFF4">
<label>4</label>
<addr-line> Department of Comparative Biomedical Sciences, University of Teramo Teramo Italy</addr-line>
</aff>
<author-notes>
<corresp id="FN1">
<label>*</label>
Corresponding author:
<email>lvesna@polj.uns.ac.rs</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub">
<year>2013</year>
</pub-date>
<pub-date pub-type="epub">
<day>14</day>
<month>1</month>
<year>2013</year>
</pub-date>
<volume>20</volume>
<issue-id pub-id-type="publisher-id">parasite/2013/01</issue-id>
<elocation-id>3</elocation-id>
<history>
<date date-type="received">
<day>18</day>
<month>9</month>
<year>2012</year>
</date>
<date date-type="accepted">
<day>26</day>
<month>11</month>
<year>2012</year>
</date>
</history>
<permissions>
<copyright-statement>© V. Lalošević et al., published by EDP Sciences, 2013</copyright-statement>
<copyright-year>2013</copyright-year>
<copyright-holder>V. Lalošević et al.,</copyright-holder>
<license license-type="open-access">
<license-p>This is an Open Access article distributed under the terms of the Creative Commons Attribution License (
<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/2.0">http://creativecommons.org/licenses/by/2.0</ext-link>
), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
</license>
</permissions>
<self-uri content-type="zoobank" xlink:type="simple" xlink:href="http://zoobank.org/urn:lsid:zoobank.org:pub:FC2CF05E-E46F-4E74-9E50-81C35EAE8E8C">urn:lsid:zoobank.org:pub:FC2CF05E-E46F-4E74-9E50-81C35EAE8E8C</self-uri>
<abstract>
<p>The respiratory capillariid nematode
<italic>Eucoleus aerophilus</italic>
(Creplin, 1839) infects wild and domestic carnivores and, occasionally, humans. Thus far, a dozen of human infections have been published in the literature but it cannot be ruled out that lung capillariosis is underdiagnosed in human medicine. Also, the apparent spreading of
<italic>E. aerophilus</italic>
in different geographic areas spurs new studies on the epidemiology of this nematode. After the recognition of the first human case of
<italic>E. aerophilus</italic>
infection in Serbia, there is a significant merit in enhancing knowledge on the distribution of the nematode. In the present work the infection rate of pulmonary capillariosis was investigated in 70 red foxes (
<italic>Vulpes vulpes</italic>
) from the northern part of Serbia by autopsy. The estimated infection rate with
<italic>Eucoleus aerophilus</italic>
was 84%. In contrast, by copromicroscopic examination only 38% of foxes were positive. In addition, 10 foxes were investigated for the closely related species in nasal cavity,
<italic>Eucoleus boehmi</italic>
, and nine were positive. Our study demonstrates one of the highest infection rates of pulmonary capillariosis in foxes over the world.</p>
</abstract>
<trans-abstract xml:lang="fr">
<p>Le nématode Capillariidae de l’appareil respiratoire
<italic>Eucoleus aerophilus</italic>
(Creplin, 1839) infecte les carnivores sauvages et domestiques et parfois les humains. À ce jour une douzaine d’infections humaines ont été publiées dans la littérature, mais il ne peut pas être exclu que la capillariose du poumon soit sous-diagnostiquée en médecine humaine. En outre, l’apparente propagation de
<italic>E. aerophilus</italic>
dans différentes zones géographiques stimule de nouvelles études sur l’épidémiologie de ce nématode. Après la reconnaissance du premier cas humain d’infection à
<italic>E. aerophilus</italic>
en Serbie, il est important d’améliorer les connaissances sur la distribution de ce nématode. Dans le présent travail le taux d’infection de capillariose pulmonaire a été étudié par autopsie chez 70 renards roux (
<italic>Vulpes vulpes</italic>
) du nord de la Serbie. Le taux d’infection à
<italic>E. aerophilus</italic>
estimé était de 84 %. En revanche, seulement 38 % des renards étaient positifs par examen copromicroscopique. En outre, dix renards ont été étudiés pour l’espèce étroitement apparentée des fosses nasales
<italic>Eucoleus boehmi</italic>
, et neuf ont été positifs. Notre étude démontre l’un des taux d’infection le plus élevés au monde de capillariose pulmonaire chez les renards.</p>
</trans-abstract>
<kwd-group>
<kwd>
<italic>Eucoleus aerophilus</italic>
</kwd>
<kwd>
<italic>Eucoleus boehmi</italic>
</kwd>
<kwd>red fox</kwd>
<kwd>Serbia</kwd>
</kwd-group>
<counts>
<fig-count count="4"></fig-count>
<table-count count="1"></table-count>
<equation-count count="0"></equation-count>
<ref-count count="33"></ref-count>
<page-count count="5"></page-count>
</counts>
</article-meta>
</front>
<body>
<sec id="S1">
<title>Introduction</title>
<p>Nematodes of the family Capillariidae Neveu-Lemaire, 1936 are a very large group with not yet completely accepted taxonomy [
<xref ref-type="bibr" rid="R16">16</xref>
,
<xref ref-type="bibr" rid="R32">32</xref>
]. Among species ranked in this family,
<italic>Eucoleus aerophilus</italic>
(Creplin, 1839) Dujardin, 1845 (syn.
<italic>Capillaria aerophila</italic>
or
<italic>Thominx aerophilus</italic>
) affects trachea and main bronchi of canids, felids and some omnivorous animals [
<xref ref-type="bibr" rid="R19">19</xref>
]. Although several morphological descriptions have been provided for different biological stages of
<italic>E. aerophilus</italic>
[
<xref ref-type="bibr" rid="R3">3</xref>
,
<xref ref-type="bibr" rid="R5">5</xref>
,
<xref ref-type="bibr" rid="R18">18</xref>
,
<xref ref-type="bibr" rid="R23">23</xref>
,
<xref ref-type="bibr" rid="R32">32</xref>
], several other aspects of this nematode are still poorly known. Indeed, this is the case of geographic distribution, clinical significance and actual zoonotic potential [
<xref ref-type="bibr" rid="R31">31</xref>
]. Also, knowledge on the biological cycle of this nematode is scanty and, for instance, the route of transmission to vertebrate hosts remains to be understood. In general it is thought that animals become infected by ingesting environmental larvated eggs [
<xref ref-type="bibr" rid="R31">31</xref>
] but it has been also hypothesized, but never demonstrated, that larval
<italic>E. aerophilus</italic>
might require the passage through earthworms to become infective for cats and foxes (Borovkova, 1947 cited in [
<xref ref-type="bibr" rid="R18">18</xref>
]).</p>
<p>In the past few years some evidences have suggested that
<italic>E. aerophilus</italic>
is spreading in several geographic areas and that there is an increase of cross-infections between wild and domestic animals [
<xref ref-type="bibr" rid="R7">7</xref>
<xref ref-type="bibr" rid="R9">9</xref>
,
<xref ref-type="bibr" rid="R30">30</xref>
,
<xref ref-type="bibr" rid="R31">31</xref>
]. Indeed, it has been preliminarily shown that some
<italic>E. aerophilus</italic>
populations are shared between domestic pets and wildlife, e.g. red fox (
<italic>Vulpes vulpes</italic>
) and beech-marten (
<italic>Martes foina</italic>
(Erxleben) [
<xref ref-type="bibr" rid="R9">9</xref>
]).</p>
<p>Such changes are a likely effect of the increase of fox populations in peri-urban and urban areas and of movements of wild and companion animals around regions. In fact, an increase of the red fox population has been documented in Europe after the success of the oral vaccination against rabies [
<xref ref-type="bibr" rid="R4">4</xref>
,
<xref ref-type="bibr" rid="R33">33</xref>
]. The role of the red fox as reservoir for parasites may be now of relevance especially in suburban areas, e.g. for multilocular echinococcosis and other parasitic diseases of veterinary (e.g. angiostrongylosis) and human (e.g. trichinellosis) concern [
<xref ref-type="bibr" rid="R11">11</xref>
,
<xref ref-type="bibr" rid="R15">15</xref>
,
<xref ref-type="bibr" rid="R21">21</xref>
,
<xref ref-type="bibr" rid="R22">22</xref>
,
<xref ref-type="bibr" rid="R31">31</xref>
].</p>
<p>With regard to Serbia,
<italic>E. aerophilus</italic>
is circulating in cats and foxes [
<xref ref-type="bibr" rid="R12">12</xref>
,
<xref ref-type="bibr" rid="R13">13</xref>
] and, interestingly, the last case of human capillariosis documented in the literature is from the same country. More specifically, an infected woman showed respiratory symptoms miming bronchial carcinoma and blood eosinophilia [
<xref ref-type="bibr" rid="R12">12</xref>
]. Considering all the aforementioned background, especially the potential risk for human health which causes new interest in capillariid nematodes, the aim of this work was to investigate the infection rate of
<italic>E. aerophilus</italic>
in red foxes of Serbia.</p>
</sec>
<sec sec-type="materials|methods" id="S2">
<title>Materials and Methods</title>
<sec id="S3">
<title>Study area</title>
<p>In winter season from 2008 to the end of 2011, foxes (
<italic>n</italic>
 = 118) hunted in the framework of a rabies control campaign were collected from the whole territory (21,506 square km) of the Vojvodina province of Serbia (45°15 N, 19°50′ E). This province is crossed by the Danube river, two tributaries (Tisza and Sava) and many canals of the Danube-Tisza-Danube system (Pannonian Basin) on the left side and the Fruska Gora mountain on the right. The climate is continental.</p>
</sec>
<sec id="S4">
<title>Sampling</title>
<p>All foxes were adults with a body weight ranging from 4.0 to 8.1 kg. Carcasses were opened and the tracheas of 70 foxes, from the larynx to the main bronchi bifurcation, were collected and preserved in 30% ethanol. Tracheas were then opened on the anterior side by scissors and parasites retrieved were collected under a stereomicroscope and wet-mounted in glycerine-ethanol or lactophenol. The nasal cavity of 10 foxes was opened through the palate, and the nasal mucosa was scrapped and conserved in 30% ethanol. Faecal samples were collected during the necropsy of the 118 foxes and tested by the glycerine flotation method, respectively. Parasites collected were counted, photographed by a Leica microscope and measured by the “ImageJ” free program (
<ext-link ext-link-type="uri" xlink:href="http://imagej.nih.gov/ij/">http://imagej.nih.gov/ij/</ext-link>
). Adult stages of nematodes recovered from trachea and bronchi of the foxes were identified at the species level by morphological keys [
<xref ref-type="bibr" rid="R17">17</xref>
,
<xref ref-type="bibr" rid="R23">23</xref>
]. Five adult female and three male
<italic>E. aerophilus</italic>
voucher specimens were deposited in the Zoological collection of the Department of Biology and Ecology, University of Novi Sad, Serbia (Registration No. NEM-0037).</p>
</sec>
</sec>
<sec sec-type="results" id="S5">
<title>Results</title>
<p>Adults of
<italic>E. aerophilus</italic>
(i.e. total of 817) were detected in 59 foxes (84%), with an average worm burden of 14 (range 1–71) (
<xref ref-type="fig" rid="F1">Figures 1</xref>
and
<xref ref-type="fig" rid="F2">2</xref>
). Out of 70 examined foxes, 32% harboured more than 14 adult worms of
<italic>E. aerophilus</italic>
with no relevant difference in parasitic burden between foxes collected from different mountainous regions or the Pannonian basin. The sex was assessed for 390 out of 817 examined worms because some
<italic>E. aerophilus</italic>
specimens were damaged during the collection. The sex ratio between male and female worms was 1:2.56.
<fig id="F1" orientation="portrait" position="float">
<label>Figure 1.</label>
<caption>
<p>Adult female
<italic>Eucoleus aerophilus</italic>
on the mucosa of the opened trachea of a fox dissected after formalin fixation. More than half of the worm body is inside the tracheal mucosa with a zig-zag shape and the other part is free in the lumen. Bar, 1 mm.</p>
</caption>
<graphic xlink:href="parasite-20-3-fig1"></graphic>
</fig>
<fig id="F2" orientation="portrait" position="float">
<label>Figure 2.</label>
<caption>
<p>Number of
<italic>Eucoleus aerophilus</italic>
adults per foxes.</p>
</caption>
<graphic xlink:href="parasite-20-3-fig2"></graphic>
</fig>
</p>
<p>Morphometric measures were obtained from 82 males and 75 females. Measurements are in micrometer (μm) and indicated here as mean and range between parentheses. Body length, male 19,020 (10,410–25,110), female 27,980 (16,010–41,840); mean diameter measured after end of oesophagus, male 86 (69–110), female 147 (107–185); oesophagus length, male 6,590 (4,390–7,980), female 6,003 (4,540–7,330); number of stichocytes, calculated in 10 males and 11 females, male 44 (37–46), female 44.5 (35–50). The lateral bacillary bands were well recognized but the ventral bacillary bands were observed rarely. The uterus containing numerous eggs continued in a muscular vagina with a vulvas opening (
<xref ref-type="fig" rid="F3">Figure 3</xref>
). Eggs were unembryonated, thick-shelled with a reticulated surface and with polar plug-like opercula, giving them a lemon shape. Eggs not symmetrical, opercular axes slightly angulated. Measurements of eggs in the final part of the uterus of 75 females: 73 × 35 (61–95 × 30–47). The caudal end of males showed a small pseudobursa with two bulges next to the cloaca opening. Males showed a single very thin spicule and a spicule sheath with many spines (
<xref ref-type="fig" rid="F4">Figure 4</xref>
).
<fig id="F3" orientation="portrait" position="float">
<label>Figure 3.</label>
<caption>
<p>Female
<italic>Eucoleus aerophilus</italic>
showing the end part of the last stichocyte with the cell nucleus (N), two superimposed secretory cells (S), intestine (I), vagina with eggs (E) and vulva (arrowhead). Bar, 50 μm.</p>
</caption>
<graphic xlink:href="parasite-20-3-fig3"></graphic>
</fig>
<fig id="F4" orientation="portrait" position="float">
<label>Figure 4.</label>
<caption>
<p>Male
<italic>Eucoleus aerophilus</italic>
, extruded spinose spicule sheath and partially extruded spicule at the caudal part. Bar, 100 μm.</p>
</caption>
<graphic xlink:href="parasite-20-3-fig4"></graphic>
</fig>
</p>
<p>For 10 foxes the nasal cavity mucosa was scrapped and examined under a stereomicroscope. Adult stages belonging to the closely related species
<italic>Eucoleus boehmi</italic>
(Supperer, 1953) were found in nine animals. The average worm burden was seven (range 1–20).</p>
<p>The results of the copromicroscopic examinations on faecal samples collected at the necropsy are reported in
<xref ref-type="table" rid="T1">Table 1</xref>
.
<table-wrap id="T1" orientation="portrait" position="float">
<label>Table 1.</label>
<caption>
<p>Results for the copromicroscopic examinations of the foxes (
<italic>n</italic>
 = 118) examined in the present study.
<italic>N</italic>
: number of foxes scored positive for endoparasites.</p>
</caption>
<table frame="hsides" rules="groups">
<thead>
<tr valign="top">
<th align="left" rowspan="1" colspan="1">Parasite species</th>
<th align="center" rowspan="1" colspan="1">
<italic>N</italic>
</th>
<th align="center" rowspan="1" colspan="1">Infection rate (%)</th>
</tr>
</thead>
<tbody>
<tr valign="top">
<td align="left" rowspan="1" colspan="1">
<italic>Toxascaris leonina</italic>
</td>
<td align="char" char="." rowspan="1" colspan="1">44</td>
<td align="char" char="." rowspan="1" colspan="1">37</td>
</tr>
<tr valign="top">
<td align="left" rowspan="1" colspan="1">
<italic>Toxocara canis</italic>
</td>
<td align="char" char="." rowspan="1" colspan="1">19</td>
<td align="char" char="." rowspan="1" colspan="1">16</td>
</tr>
<tr valign="top">
<td align="left" rowspan="1" colspan="1">
<italic>Trichuris vulpis</italic>
</td>
<td align="char" char="." rowspan="1" colspan="1">13</td>
<td align="char" char="." rowspan="1" colspan="1">11</td>
</tr>
<tr valign="top">
<td align="left" rowspan="1" colspan="1">
<italic>Ancylostoma</italic>
spp.</td>
<td align="char" char="." rowspan="1" colspan="1">29</td>
<td align="char" char="." rowspan="1" colspan="1">25</td>
</tr>
<tr valign="top">
<td align="left" rowspan="1" colspan="1">Capillariidae gen
<italic>.</italic>
spp.</td>
<td align="char" char="." rowspan="1" colspan="1">45</td>
<td align="char" char="." rowspan="1" colspan="1">38</td>
</tr>
<tr valign="top">
<td align="left" rowspan="1" colspan="1">
<italic>Isospora</italic>
spp.</td>
<td align="char" char="." rowspan="1" colspan="1">1</td>
<td align="char" char="." rowspan="1" colspan="1">0.84</td>
</tr>
<tr valign="top">
<td align="left" rowspan="1" colspan="1">Parasite free</td>
<td align="char" char="." rowspan="1" colspan="1">23</td>
<td align="char" char="." rowspan="1" colspan="1">20</td>
</tr>
</tbody>
</table>
</table-wrap>
</p>
</sec>
<sec sec-type="discussion" id="S6">
<title>Discussion</title>
<p>The infection rate of
<italic>E. aerophilus</italic>
infection (84%) herein detected in foxes from the Pannonian and the Fruska Gora Mountain regions (Vojvodina province) is indeed high. Interestingly, along with the results (88%) recently obtained from foxes of Norway [
<xref ref-type="bibr" rid="R6">6</xref>
], this study demonstrated the highest infection rate of pulmonary capillariosis in wild canids in Europe.</p>
<p>In the Zagreb region (Croatia) which shows a similar landscape to the central part of Serbia, about 300 km on the West, the prevalence of
<italic>E. aerophilus</italic>
in foxes was only 4.7% with a highest worm burden of five worms [
<xref ref-type="bibr" rid="R20">20</xref>
]. In Italy, the infection rate of
<italic>E. aerophilus</italic>
detected in the trachea of foxes was 7.0% in the Tuscany region [
<xref ref-type="bibr" rid="R14">14</xref>
]. In the Netherlands and Hungary
<italic>E. aerophilus</italic>
was found in 46.8% and 66% of dissected foxes [
<xref ref-type="bibr" rid="R1">1</xref>
,
<xref ref-type="bibr" rid="R27">27</xref>
]. An infection rate by “
<italic>Capillaria</italic>
spp.” of 22.4% was found in foxes from Slovakia by copromicroscopic investigation [
<xref ref-type="bibr" rid="R15">15</xref>
].</p>
<p>Albeit the high prevalence of
<italic>E. aerophilus</italic>
in foxes from Serbia could be due to the high sensitivity of the diagnostic methods used, most of the aforementioned studies have relied on trachea dissection as well. A great discrepancy between copromicroscopic and necroscopical findings of capillariids in the present study demonstrates that copromicroscopy may fail in terms of diagnostic sensitivity for this infection in foxes. Therefore, factors other than the diagnostic approach should be implicated in influencing such a high infection rate by lung capillariosis in foxes from Serbia.</p>
<p>The high humidity of the habitat of Vojvodina, the South part of Pannonian Basin, with many canals between the Danube and Tisza rivers, and many small tributaries of the Sava river in the Fruska Gora Mountain could favour parasite’s dissemination. In this habitat, the fox density is of over 0.2 per square km and the high humidity can favour earthworms’ dispersion. Also, competitive pressure and other diseases (e.g. sarcoptic mange) can enhance the susceptibility of foxes to nematodes [
<xref ref-type="bibr" rid="R24">24</xref>
] and the increase of reports of capillariosis in companion animals suggests that
<italic>E. aerophilus</italic>
is spreading in several areas. Epidemiological data (e.g. range of hosts and geographic distribution) of
<italic>E. aerophilus</italic>
in Europe are poorly known, thus at the moment it is difficult to assess to what degree this parasite may be spreading or what influence global warming or other factors may have on the current distribution [
<xref ref-type="bibr" rid="R31">31</xref>
]. On the other hand, it is known that wild foxes act as reservoirs and amplifiers of several canid nematodes, thus they can re-enforce environmental contamination and risk of infection for domestic dogs and humans as well [
<xref ref-type="bibr" rid="R29">29</xref>
].</p>
<p>There is still not an agreement of the taxonomy of capillariid nematodes. Key morphological characters (i.e. eggs, oesophageal structure and genital organs of adult stages) allow the identification of
<italic>E. aerophilus</italic>
but, sometimes its identification at the species level and the differentiation from
<italic>E. boehmi</italic>
may be difficult.</p>
<p>While Christenson [
<xref ref-type="bibr" rid="R5">5</xref>
] did not detect spicule and spicule sheath in male
<italic>E. aerophilus</italic>
, more recent line drawings of this very characteristic organ were published by Butterworth and Beverley-Burton [
<xref ref-type="bibr" rid="R3">3</xref>
], Moravec [
<xref ref-type="bibr" rid="R17">17</xref>
] and Romashov [
<xref ref-type="bibr" rid="R23">23</xref>
]. The present work confirms the presence of these morphological features, which is of key diagnostic relevance for the identification at the species level of male
<italic>E. aerophilus</italic>
.</p>
<p>The stichocyte number, higher in
<italic>E. aerophilus</italic>
than in
<italic>E. boehmi</italic>
, can help to distinguish these two species between them, even though in some cases, the number of stichocytes may overlap. In fact in the present study, one
<italic>E. boehmi</italic>
female showed 35 stichocytes as an
<italic>E. aerophilus</italic>
female. According to Moravec [
<xref ref-type="bibr" rid="R17">17</xref>
],
<italic>E. boehmi</italic>
and
<italic>E. aerophilus</italic>
females have 30–32 and 42–46 stichocytes, respectively, whereas, according to Romashov [
<xref ref-type="bibr" rid="R23">23</xref>
], the number of stichocytes in the female is 32–36 for
<italic>E. boehmi</italic>
and 35–49 for
<italic>E. aerophilus</italic>
. Greater variations of the stichocyte number were reported in
<italic>E. aerophilus</italic>
males, i.e. 42–55 [
<xref ref-type="bibr" rid="R23">23</xref>
], or 43–50 [
<xref ref-type="bibr" rid="R17">17</xref>
]. The present study revealed that the number of stichocytes may be useful in the identification of
<italic>Eucoleus</italic>
spp. but it is not absolute criteria. The localization of adult stages could also greatly help in distinguishing between
<italic>E. aerophilus</italic>
(in trachea and bronchi) and
<italic>E. boehmi</italic>
(in nasal cavities).</p>
<p>Another issue warranting further investigations is the size of the eggs. While some textbooks report a length of up to 83 μm for
<italic>E. aerophilus</italic>
eggs, e.g. Soulsby [
<xref ref-type="bibr" rid="R26">26</xref>
], Bowman
<italic>et al.</italic>
[
<xref ref-type="bibr" rid="R2">2</xref>
], Taylor
<italic>et al.</italic>
[
<xref ref-type="bibr" rid="R28">28</xref>
], recent studies have demonstrated that the length of
<italic>E. aerophilus</italic>
eggs from dogs and cats is unwaveringly less than 70 μm [
<xref ref-type="bibr" rid="R7">7</xref>
,
<xref ref-type="bibr" rid="R8">8</xref>
,
<xref ref-type="bibr" rid="R32">32</xref>
]. Interestingly, some ova collected from the uterus of adult
<italic>E. aerophilus</italic>
in the present study were found to be longer and this could be from different reasons. It could be that eggs undergo further development and modifications when they are passed with the faeces, or the existence of different morphotypes within the same species
<italic>E. aerophilus</italic>
and with different host affiliations could be hypothesized. Recent molecular studies have shown that different haplotypes of
<italic>E. aerophilus</italic>
may be either shared between domestic and wild carnivores or are affiliated to specific hosts [
<xref ref-type="bibr" rid="R9">9</xref>
]. Further studies are necessary for evaluating the distribution of
<italic>E. aerophilus</italic>
in wildlife and pets cohabiting the same geographic areas in order to elucidate the phylogeography of different parasite populations and for verifying the possible existence of different morphotypes [
<xref ref-type="bibr" rid="R8">8</xref>
,
<xref ref-type="bibr" rid="R9">9</xref>
].</p>
<p>Under a clinical standpoint, the pathogenic role of
<italic>E. aerophilus</italic>
in wildlife is not well recognized because it is ranging from low to high and
<italic>E. aerophilus</italic>
has been also considered as an agent of massive mortality in farmed silver foxes [
<xref ref-type="bibr" rid="R25">25</xref>
]. In the present work, a very low inflammatory response was found around the worms (data not shown) which can be the result of a low pathogenicity or an immunosuppression in the mucosa caused by the parasite in these animals. A past work carried out in six experimentally infected foxes showed that animals may develop bronchopneumonia with cough and other signs of heavy respiratory infection, till death in some cases (Borovkova, cited in [
<xref ref-type="bibr" rid="R25">25</xref>
]). In pets, this parasite may cause bronchovesicular sounds, respiratory inflammation, sneezing, wheezing, chronic moist or dry cough, (broncho)-pneumonia respiratory failure and heavy parasite burdens may lead to mortality [
<xref ref-type="bibr" rid="R10">10</xref>
,
<xref ref-type="bibr" rid="R31">31</xref>
]. More studies are also necessary to evaluate the actual role of
<italic>E. aerophilus</italic>
in causing undiagnosed respiratory distresses in companion dogs and cats.</p>
<p>Another topic which needs to be better elucidated is the actual role of
<italic>E. aerophilus</italic>
in causing lung diseases in humans. Indeed, only 12 cases of infection have been published in the literature [
<xref ref-type="bibr" rid="R13">13</xref>
], but it is likely that the infection is underdiagnosed because the clinical signs may overlap a plethora of respiratory diseases which may be self-limiting or may resolve after non-specific treatments.</p>
</sec>
</body>
<back>
<ack>
<p>Authors are thankful to Dr. Edoardo Pozio, Istituto Superiore di Sanità, Rome, for critical reading of the manuscript. This study was supported by Ministry of Education and Science of Serbia, Grant No. TR31084.</p>
</ack>
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<bold>Cite this article as</bold>
: Lalošević V, Laloševicć D, Čapo I, Simin V, Galfi A & Traversa D: High infection rate of zoonotic
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